Carbohydrate-chain analysis by lectin binding to mixtures of glycoproteins, separated by polyacrylamide slab-gel electrophoresis, with in situ chemical modifications
✍ Scribed by Tatsuro Irimura; Garth L. Nicolson
- Publisher
- Elsevier Science
- Year
- 1983
- Tongue
- English
- Weight
- 717 KB
- Volume
- 115
- Category
- Article
- ISSN
- 0008-6215
No coin nor oath required. For personal study only.
✦ Synopsis
The interactions of five 1251-lectins of differing binding-specificities with seven glycoproteins containing known carbohydrate chains were investigated on polyacrylamide gels after electrophoretic separation. The glycoproteins were also chemically modified in the gels in order to gather additional information, The glycoproteins used were: porcine thyroglobulin (PTG); bovine-serum fibronectin (BFN); humanserum transferrin (HTF), bovine fetuin (BFE); subfractionated, hen ovalbumins having galactosylated, hybrid-type, carbohydrate chains (OVG); agalactosyl, hybridtype-carbohydrate chains (OVH); and high-mannose-type-carbohydrate chains (OVM). Wheat-germ agglutinin (WGA) stained BFE, OVG, and OVH, but removal of sialic acid from BFE resulted in loss of WGA binding. After desialosylation and Smith degradation in situ, BFE and HTF reacted strongly, and PTG slightly with WGA. Ricinus conmunis agglutinin I stained PTG, BFN, HTF, and BFE after removal of sialic acid. Arachis hypogaea (peanut) agglutinin reacted only with BFN after removal of sialic acid. Lens culinaris hemagglutinin bound only to PTG, and concanavalin A bound to PTG, BFE, HTF, OVH, and OVM. The results were consistent with previously proposed lectin-carbohydrate-binding specificities, indicating that lectins may be used to determine the carbohydrate-chain structures of particular glycoproteins, even in complex mixtures.